TY - JOUR
T1 - Protein purification, crystallization and preliminary X-ray diffraction analysis of l-arabinose isomerase from Lactobacillus fermentum CGMCC2921
AU - Xu, Zheng
AU - Li, Sha
AU - Liang, Jinfeng
AU - Feng, Xiaohai
AU - Xu, Hong
N1 - Publisher Copyright:
© 2015 International Union of Crystallography.
PY - 2015/1/1
Y1 - 2015/1/1
N2 - l-Arabinose isomerase (AI) catalyzes the isomerization of l-arabinose to l-ribulose, as well as that of d-galactose to d-tagatose. A thermophilic AI derived from Lactobacillus fermentum CGMCC2921 (LFAI) was overexpressed in Escherichia coli BL21 (DE3). This enzyme was purified to over 95% purity by nickel affinity, Mono-Q ion-exchange and size-exclusion chromatography. The LFAI protein was crystallized from either 0.1M bis-tris pH 6.5, 23% PEG 3350, 0.3M NaCl (form 1 crystals) or 0.1M bis-tris pH 6.0, 25% PEG monomethyl ether 5000 (form 2 crystals). Diffraction data from form 1 LFAI crystals were collected to 2.80Å resolution using synchrotron radiation. The form 1 crystals belonged to the orthorhombic space group P212121, with unit-cell parameters a = 85.11, b = 184.57, c = 186.26Å, α = β = γ = 90°. The asymmetric unit contained six LFAI subunits, corresponding to a calculated Matthews coefficient of 2.29Å3Da-1 and a solvent content of 46.22%.
AB - l-Arabinose isomerase (AI) catalyzes the isomerization of l-arabinose to l-ribulose, as well as that of d-galactose to d-tagatose. A thermophilic AI derived from Lactobacillus fermentum CGMCC2921 (LFAI) was overexpressed in Escherichia coli BL21 (DE3). This enzyme was purified to over 95% purity by nickel affinity, Mono-Q ion-exchange and size-exclusion chromatography. The LFAI protein was crystallized from either 0.1M bis-tris pH 6.5, 23% PEG 3350, 0.3M NaCl (form 1 crystals) or 0.1M bis-tris pH 6.0, 25% PEG monomethyl ether 5000 (form 2 crystals). Diffraction data from form 1 LFAI crystals were collected to 2.80Å resolution using synchrotron radiation. The form 1 crystals belonged to the orthorhombic space group P212121, with unit-cell parameters a = 85.11, b = 184.57, c = 186.26Å, α = β = γ = 90°. The asymmetric unit contained six LFAI subunits, corresponding to a calculated Matthews coefficient of 2.29Å3Da-1 and a solvent content of 46.22%.
KW - d-tagatose
KW - l-arabinose isomerase
KW - rare sugars
KW - synchrotron radiation
UR - http://www.scopus.com/inward/record.url?scp=84921775908&partnerID=8YFLogxK
U2 - 10.1107/S2053230X14025321
DO - 10.1107/S2053230X14025321
M3 - 文章
C2 - 25615964
AN - SCOPUS:84921775908
SN - 1744-3091
VL - 71
SP - 28
EP - 33
JO - Acta Crystallographica Section F:Structural Biology Communications
JF - Acta Crystallographica Section F:Structural Biology Communications
ER -